scholarly journals Phenotypic detection and genotyping of Clostridium perfringens associated with enterotoxemia in sheep in the Qassim Region of Saudi Arabia

2021 ◽  
Vol 14 (3) ◽  
pp. 578-584
Author(s):  
Fehaid Alsaab ◽  
Ali Wahdan ◽  
Elhassan M. A. Saeed
Keyword(s):  
Saudi Arabia ◽  
Clostridium Perfringens ◽  
Culture Method ◽  
Economic Loss ◽  
Intestinal Content ◽  
Toxin Gene ◽  
Rt Pcr ◽  
Toxin Genes ◽  
Type D ◽  
Vitek 2

Background and Aim: Enterotoxemia caused by Clostridium perfringens toxinotypes is an often fatal disease of sheep of all ages, with a substantial economic loss to the sheep industry. This study was conducted to isolate C. perfringens from suspected cases of enterotoxemia in sheep in the central part of the Qassim Region, Saudi Arabia, and to determine the prevalent toxinotype by detecting alpha (cpA), beta (cpB), and epsilon (etX) toxin genes, which might help control this disease locally. Materials and Methods: A total of 93 rectal swabs and intestinal content samples were collected from diseased and animals suspected of having died of enterotoxemia in early 2020. Samples were subjected to bacteriological examination, biochemical analysis of isolates by VITEK 2, and molecular toxinotyping of isolates by LightCycler® real-time polymerase chain reaction (RT-PCR). Results: Our results revealed that only 14 isolates were confirmed by VITEK 2 as being C. perfringens, with excellent identification (probability of 95% and 97%). According to the toxinotyping of isolates by RT-PCR, all 14 isolates possessed both the cpA and etX toxin genes, while the cpB toxin gene was not detected in any of the isolates. Conclusion: Our findings demonstrated that C. perfringens type D was the only toxinotype found in the central part of the Qassim Region in 2020; moreover, according to the culture method, only 15% (14/93) of the suspected cases of enterotoxemia were confirmed to be caused by C. perfringens infection, which highlighted the importance of clinical and laboratory differential diagnosis of enterotoxemia in sheep.

scholarly journals Genomic Analysis of Clostridium perfringens BEC/CPILE-Positive, Toxinotype D and E Strains Isolated from Healthy Children

Toxins ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 543
Author(s):  
Raymond Kiu ◽  
Kathleen Sim ◽  
Alex Shaw ◽  
Emma Cornwell ◽  
Derek Pickard ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Genomic Analysis ◽  
Future Research ◽  
Toxin Gene ◽  
Healthy Children ◽  
Healthy Individuals ◽  
Type D ◽  
Plasmid Analysis ◽  
Putative Toxin ◽  
Specific Trait

Clostridium perfringens toxinotype D, toxinotype E, and gastroenteritis-linked BEC/CPILE-positive strains have never been reported in healthy children. We isolated, whole-genome sequenced and bioinformatically characterised three C. perfringens isolates—type D (IQ1), type E (IQ2) and BEC/CPILE-positive (IQ3), recovered from the stools of three healthy two-year-olds, which were further compared to 128 C. perfringens genomes available from NCBI. The analysis uncovered a previously under-described putative toxin gene alv (alveolysin) encoded by isolates IQ2 and IQ3, which appeared to be a clade-specific trait associated with strains from domestic animals. A plasmid analysis indicated that the iota-toxin was encoded on a near-intact previously described plasmid pCPPB-1 in type E strain IQ2. The BEC genes becA and becB were carried on a near-identical pCPOS-1 plasmid previously associated with Japanese gastroenteritis outbreaks. Furthermore, a close phylogenetic relatedness was inferred between the French C. perfringens type E isolates cp515.17 and newly sequenced IQ2, suggesting geographical links. This study describes novel C. perfringens isolates from healthy individuals which encode important toxin genes, indicating the potential spread of these veterinary and clinically important strains and mobile genetic elements, and highlights areas for future research.

scholarly journals Sequencing and Diversity Analyses Reveal Extensive Similarities between Some Epsilon-Toxin-Encoding Plasmids and the pCPF5603 Clostridium perfringens Enterotoxin Plasmid

2008 ◽  
Vol 190 (21) ◽  
pp. 7178-7188 ◽  
Author(s):  
Kazuaki Miyamoto ◽  
Jihong Li ◽  
Sameera Sayeed ◽  
Shigeru Akimoto ◽  
Bruce A. McClane
Keyword(s):  
Clostridium Perfringens ◽  
Open Reading Frames ◽  
Toxin Gene ◽  
Type B ◽  
Clostridium Perfringens Enterotoxin ◽  
Type D ◽  
Transfer Genes ◽  
Epsilon Toxin ◽  
Beta2 Toxin ◽  
Beta Toxin

ABSTRACT Clostridium perfringens type B and D isolates produce epsilon-toxin, the third most potent clostridial toxin. The epsilon-toxin gene (etx) is plasmid borne in type D isolates, but etx genetics have been poorly studied in type B isolates. This study reports the first sequencing of any etx plasmid, i.e., pCP8533etx, from type B strain NCTC8533. This etx plasmid is 64.7 kb, carries tcp conjugative transfer genes, and encodes additional potential virulence factors including beta2-toxin, sortase, and collagen adhesin but not beta-toxin. Interestingly, nearly 80% of pCP8533etx open reading frames (ORFs) are also present on pCPF5603, an enterotoxin-encoding plasmid from type A isolate F5603. Pulsed-field gel electrophoresis and overlapping PCR indicated that a pCP8533etx-like etx plasmid is also present in most, if not all, other type B isolates and some beta2-toxin-positive, cpe-negative type D isolates, while other type D isolates carry different etx plasmids. Sequences upstream of the etx gene vary between type B isolates and some type D isolates that do not carry a pCP8533etx-like etx plasmid. However, nearly all type B and D isolates have an etx locus with an upstream IS1151, and those etx loci typically reside near a dcm ORF. These results suggest that pCPF5603 and pCP8533etx evolved from insertion of mobile genetic elements carrying enterotoxin or etx genes, respectively, onto a common progenitor plasmid.

scholarly journals Characterization of Clostridium perfringens TpeL Toxin Gene Carriage, Production, Cytotoxic Contributions, and Trypsin Sensitivity

2015 ◽  
Vol 83 (6) ◽  
pp. 2369-2381 ◽  
Author(s):  
Jianming Chen ◽  
Bruce A. McClane
Keyword(s):  
Clostridium Perfringens ◽  
Toxin Gene ◽  
Type B ◽  
Content Type ◽  
Positive Type ◽  
Natural Production ◽  
Type D ◽  
Clostridial Species ◽  
Beta Toxin

Large clostridial toxins (LCTs) are produced by at least four pathogenic clostridial species, and several LCTs are proven pivotal virulence factors for both human and veterinary diseases. TpeL is a recently identified LCT produced byClostridium perfringensthat has received relatively limited study. In response, the current study surveyed carriage of thetpeLgene among differentC. perfringensstrains, detecting this toxin gene in some type A, B, and C strains but not in any type D or E strains. This study also determined that all tested strains maximally produce, and extracellularly release, TpeL at the late-log or early-stationary growth stage duringin vitroculture, which is different from the maximal late-stationary-phase production reported previously for other LCTs and for TpeL production byC. perfringensstrain JIR12688. In addition, the present study found that TpeL levels in culture supernatants can be repressed by either glucose or sucrose. It was also shown that, at natural production levels, TpeL is a significant contributor to the cytotoxic activity of supernatants from cultures oftpeL-positive strain CN3685. Lastly, this study identified TpeL, which presumably is produced in the intestines during diseases caused by TpeL-positive type B and C strains, as a toxin whose cytotoxicity decreases after treatment with trypsin; this finding may have pathophysiologic relevance by suggesting that, like beta toxin, TpeL contributes to type B and C infections in hosts with decreased trypsin levels due to disease, diet, or age.

scholarly journals Antimicrobial Susceptibility and Association with Toxin Determinants in Clostridium perfringens Isolates from Chickens

2020 ◽  
Vol 8 (11) ◽  
pp. 1825
Author(s):  
Bai Wei ◽  
Se-Yeoun Cha ◽  
Jun-Feng Zhang ◽  
Ke Shang ◽  
Hae-Chul Park ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Clostridium Perfringens ◽  
Clinical Signs ◽  
Toxin Gene ◽  
Necrotic Enteritis ◽  
Growth Promoters ◽  
Complete Understanding ◽  
Toxin Genes ◽  
Antimicrobial Growth Promoters ◽  
Selection Of

The aim of the present study was to investigate variation in antimicrobial resistance in Clostridium perfringens (C. perfringens) isolated from chickens after withdrawal of antimicrobial growth promoters (AGPs); and to investigate the correlation between the presence of toxin genes (cpb2, netB, and tpeL) and antimicrobial resistance. Altogether, 162 isolates of C. perfringens were obtained from chickens displaying clinical signs of necrotic enteritis (n = 65) and from healthy chickens (n = 97) in Korea during 2010–2016. Compared to before AGP withdrawal, increased antimicrobial resistance or MIC50/MIC90 value was observed for nine antimicrobials including penicillin, tetracycline, tylosin, erythromycin, florfenicol, enrofloxacin, monensin, salinomycin, and maduramycin. Significantly (p < 0.05) higher resistance to gentamicin, clindamycin, and virginiamycin was found in isolates from chickens with necrotic enteritis compared to those from healthy chickens. tpeL gene was not detected in C. perfringens isolates from healthy chickens. A correlation between toxin gene prevalence and antibiotic resistance was found in the C. perfringens isolates. Because the usage of antimicrobials may contribute to the selection of both resistance and toxin genes, these can potentially make it challenging to control antimicrobial resistance in pathogenic colonies. Therefore, a more complete understanding of the interplay between resistance and virulence genes is required.

scholarly journals Characterization of Toxin Plasmids in Clostridium perfringens Type C Isolates

2010 ◽  
Vol 78 (11) ◽  
pp. 4860-4869 ◽  
Author(s):  
Abhijit Gurjar ◽  
Jihong Li ◽  
Bruce A. McClane
Keyword(s):  
Clostridium Perfringens ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Toxin Gene ◽  
Gene Encoding ◽  
Positive Type ◽  
Toxin Genes ◽  
Type C ◽  
Beta2 Toxin ◽  
Beta Toxin

ABSTRACT Clostridium perfringens type C isolates cause enteritis necroticans in humans or necrotizing enteritis and enterotoxemia in domestic animals. Type C isolates always produce alpha toxin and beta toxin but often produce additional toxins, e.g., beta2 toxin or enterotoxin. Since plasmid carriage of toxin-encoding genes has not been systematically investigated for type C isolates, the current study used Southern blot hybridization of pulsed-field gels to test whether several toxin genes are plasmid borne among a collection of type C isolates. Those analyses revealed that the surveyed type C isolates carry their beta toxin-encoding gene (cpb) on plasmids ranging in size from ∼65 to ∼110 kb. When present in these type C isolates, the beta2 toxin gene localized to plasmids distinct from the cpb plasmid. However, some enterotoxin-positive type C isolates appeared to carry their enterotoxin-encoding cpe gene on a cpb plasmid. The tpeL gene encoding the large clostridial cytotoxin was localized to the cpb plasmids of some cpe-negative type C isolates. The cpb plasmids in most surveyed isolates were found to carry both IS1151 sequences and the tcp genes, which can mediate conjugative C. perfringens plasmid transfer. A dcm gene, which is often present near C. perfringens plasmid-borne toxin genes, was identified upstream of the cpb gene in many type C isolates. Overlapping PCR analyses suggested that the toxin-encoding plasmids of the surveyed type C isolates differ from the cpe plasmids of type A isolates. These findings provide new insight into plasmids of proven or potential importance for type C virulence.

scholarly journals Complete genomic sequence and analysis of β2 toxin gene mapping of Clostridium perfringens JXJA17 isolated from piglets in China

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Xiu Zeng ◽  
Baosheng Liu ◽  
Jiao Zhou ◽  
Yimin Dai ◽  
Chunsheng Han ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Genomic Sequence ◽  
Antibiotic Resistance Genes ◽  
Close Relation ◽  
Illumina Miseq ◽  
Opportunistic Pathogen ◽  
Toxin Gene ◽  
Circular Chromosome ◽  
Toxin Genes ◽  
Genome Homology

AbstractClostridium perfringens (Cp) is a ubiquitous opportunistic pathogen of humans and animals in the natural environment and animal intestines. The pathogenicity of Cp depends on the production of toxins encoded by genes on the chromosomes or plasmids. In contemporary literature, there is no clear consensus about the pathogenicity of CpA β2 toxin. To analyze the homology of the genome of piglet source CpA and its β2 toxin, we sequenced the whole genome of strain JXJA17 isolated from diarrhea piglets using the Illumina Miseq and Pacbio Sequel platforms. The genome was composed of a circular chromosome with 3,324,072 bp (G + C content: 28.51%) and nine plasmids. Genome and 16S rDNA homology analysis revealed a close relation of the JXJA17 strain with the JGS1495, Cp-06, Cp-16, and FORC_003 strains. These strains were isolated from different samples and belonged to different toxin-types. JXJA17 strain was found to carry two toxin genes (plc and cpb2). In contrast to other Cp strains, the cpb2 of JXJA17 was located on a large plasmid (58 kb) with no co-localization of other toxin genes or antibiotic resistance genes. Analysis of JXJA17 genome homology and its cpb2 would facilitate our further study the relationship between β2 toxin and piglet diarrhea.

scholarly journals Prevalence and Genetic Diversity of Toxin Genes in Clinical Isolates of Clostridium perfringens: Coexistence of Alpha-Toxin Variant and Binary Enterotoxin Genes (bec/cpile)

Toxins ◽  
2019 ◽  
Vol 11 (6) ◽  
pp. 326 ◽  
Author(s):  
Asami Matsuda ◽  
Meiji Soe Aung ◽  
Noriko Urushibara ◽  
Mitsuyo Kawaguchiya ◽  
Ayako Sumi ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Clostridium Perfringens ◽  
Clinical Isolates ◽  
Nucleotide Sequencing ◽  
Toxin Gene ◽  
Alpha Toxin ◽  
Toxin Genes ◽  
Food Borne ◽  
Putative Origin ◽  
High Level

Clostridium perfringens (C. perfringens) is responsible for food-borne gastroenteritis and other infectious diseases, and toxins produced by this bacterium play a key role in pathogenesis. Although various toxins have been described for C. perfringens isolates from humans and animals, prevalence of individual toxins among clinical isolates has not yet been well explored. In the present study, a total of 798 C. perfringens clinical isolates were investigated for prevalence of eight toxin genes and their genetic diversity by PCR, nucleotide sequencing, and phylogenetic analysis. Besides the alpha-toxin gene (plc) present in all the isolates, the most common toxin gene was cpe (enterotoxin) (34.2%), followed by cpb2 (beta2 toxin) (1.4%), netB (NetB) (0.3%), and bec/cpile (binary enterotoxin BEC/CPILE) (0.1%), while beta-, epsilon-, and iota-toxin genes were not detected. Genetic analysis of toxin genes indicated a high level of conservation of plc, cpe, and netB. In contrast, cpb2 was revealed to be considerably divergent, containing at least two lineages. Alpha-toxin among 46 isolates was classified into ten sequence types, among which common types were distinct from those reported for avian isolates. A single isolate with bec/cpile harbored a plc variant containing an insertion of 834-bp sequence, suggesting its putative origin from chickens.

scholarly journals Genotyping of Clostridium perfringens Isolates from Domestic Livestock in Saudi Arabia

2020 ◽  
Vol 2020 ◽  
pp. 1-9 ◽  
Author(s):  
Sawsan A. Omer ◽  
Ebtesam M. Al-Olayan ◽  
Salah Eldin H. Babiker ◽  
Mohammed Z. Aljulaifi ◽  
Abdulaziz N. Alagaili ◽  
...  
Keyword(s):  
Saudi Arabia ◽  
Clostridium Perfringens ◽  
Gene Sequences ◽  
Kingdom Of Saudi Arabia ◽  
Type D ◽  
Likelihood Analysis ◽  
Genes Encoding ◽  
Domestic Livestock ◽  
Type C ◽  
Polymerase Chain

The present study was undertaken to confirm the genetic identity of Clostridium perfringens isolates from domestic livestock in Saudi Arabia and to characterize the genes encoding to alpha, beta, epsilon, and iota (α-, β-, ε-, and ι-) toxins. C. perfringens were confirmed in 104 out of 136 isolates on multiplex polymerase chain reaction using specific primers amplifying genes related to toxins produced by C. perfringens. Genes encoding α-toxins were detected in 104 samples. Of the isolates, 80.8% were diagnosed as type A, 15.4% as type D, 2.9% as type C, and 0.96% as type B. None of the isolates has genes encoding iota (ι-) toxin. All isolates investigated yielded enterotoxin (cpe) products and none yielded β2 (cpb2-toxin) or NetB products. PLC gene sequences encoding α-toxin showed >96.7% similarity. Isolates which had α-toxins as well as enterotoxin (cpe) are regarded as type F. Phylogenetic analysis using maximum likelihood analysis yielded two clades, and the majority of the isolates were in one group while only two isolates clustered on the second clade. Within the Kingdom of Saudi Arabia strains, 54 variable positions and 23 polymorphic amino acids were noticed. Isolates with ε- and β-toxins were variable and were found to be close to those published for C. perfringens. ETX gene sequences encoding ε-toxins were found to be related to CPE sequences.

Clostridium perfringens type-D enterotoxaemia in cattle: the diagnostic significance of intestinal epsilon toxin

2015 ◽  
Vol 177 (15) ◽  
pp. 390-390 ◽  
Author(s):  
A. L. Jones ◽  
M. P. Dagleish ◽  
G. L. Caldow
Keyword(s):  
Clostridium Perfringens ◽  
Case Series ◽  
Intestinal Content ◽  
Diagnostic Value ◽  
Diagnostic Significance ◽  
Type D ◽  
Intestinal Contents ◽  
Epsilon Toxin ◽  
Brain Changes ◽  
The Brain

The aims of this study were to describe 42 cases of Clostridium perfringens type-D enterotoxaemia in cattle seen between 2003 and 2014 and to determine the diagnostic value of detecting epsilon toxin in bovine intestinal content. All cases in the series had histological brain changes considered pathognomonic for C. perfringens type-D enterotoxaemia in sheep and goats and the epsilon toxin of C. perfringens was concurrently detected in the intestinal contents of 15 (36 per cent) cases. The data from the case series indicate that intestinal epsilon toxin has a sensitivity of 56 per cent compared with histology of the brain for diagnosis of bovine C. perfringens type-D enterotoxaemia. The diagnostic specificity of detecting epsilon toxin in bovine intestinal content was investigated by screening intestinal contents of 60 bovine carcases submitted for postmortem examination. Epsilon toxin was detected in 11 (18 per cent) carcases but no pathognomonic histological brain change was found in any. The specificity of intestinal epsilon toxin was estimated to be 80.4 per cent. These studies demonstrate that for a definitive diagnosis of C. perfringens type-D enterotoxaemia in cattle histological examination of the brain is essential as the presence of epsilon toxin in the intestinal contents alone is neither sensitive nor specific enough.

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